Greenspan D S, Weissman S M
Mol Cell Biol. 1985 Aug;5(8):1894-900. doi: 10.1128/mcb.5.8.1894-1900.1985.
The herpes simplex virus type 1 thymidine kinase (tk) gene lacks introns and produces stable mRNA in the absence of splicing. We have prepared a hybrid gene by placing the first exon, first intron (first intervening sequence, designated IVS1), and most of the second exon of the normal human beta-globin gene into the 3' untranslated region of the tk gene. Although this hybrid gene contains all globin sequences presumed necessary for the splicing of IVS1, predominantly, unspliced stable cytoplasmic RNA is produced in both long- and short-term expression assays. Moreover, stable unspliced cytoplasmic RNA is detected whether the intron is situated in a sense or an antisense orientation. Efficient splicing of IVS1 is obtained either by deleting the majority of tk coding sequences or by relocating the globin sequences from the 3' to the 5' untranslated region of the tk gene.
单纯疱疹病毒1型胸苷激酶（tk）基因不含内含子，在不进行剪接的情况下可产生稳定的信使核糖核酸（mRNA）。我们通过将正常人β珠蛋白基因的第一个外显子、第一个内含子（第一个间隔序列，称为IVS1）和大部分第二个外显子置于tk基因的3'非翻译区，制备了一个杂交基因。尽管这个杂交基因包含了IVS1剪接所需的所有假定珠蛋白序列，但在长期和短期表达试验中，主要产生的是未剪接的稳定细胞质RNA。此外，无论内含子以正义还是反义方向定位，都能检测到稳定的未剪接细胞质RNA。通过删除大部分tk编码序列或通过将珠蛋白序列从tk基因的3'非翻译区重新定位到5'非翻译区，可实现IVS1的有效剪接。
